Pervasive mRNA uridylation in fission yeast is catalysed by both Cid1 and Cid16 terminal uridyltransferases

Messenger RNA uridylation is pervasive and conserved among eukaryotes, but the consequences of this modification for mRNA fate are still under debate. Utilising a simple model organism to study uridylation may facilitate efforts to understand the cellular function of this process. Here we demonstrate that uridylation can be detected using simple bioinformatics approach. We utilise it to unravel widespread transcript uridylation in fission yeast and demonstrate the contribution of both Cid1 and Cid16, the only two annotated terminal uridyltransferases (TUT-ases) in this yeast. To detect uridylation in transcriptome data, we used a RNA-sequencing (RNA-seq) library preparation protocol involving initial linker ligation to fragmented RNA-an approach borrowed from small RNA sequencing that was commonly used in older RNA-seq protocols. We next explored the data to detect uridylation marks. Our analysis show that uridylation in yeast is pervasive, similarly to the one in multicellular organisms. Importantly, our results confirm the role of the cytoplasmic uridyltransferase Cid1 as the primary uridylation catalyst. However, we also observed an auxiliary role of the second uridyltransferase, Cid16. Thus both fission yeast uridyltransferases are involved in mRNA uridylation. Intriguingly, we found no physiological phenotype of the single and double deletion mutants of cid1 and cid16 and only minimal impact of uridylation on steady-state mRNA levels. Our work establishes fission yeast as a potent model to study uridylation in a simple eukaryote, and we demonstrate that it is possible to detect uridylation marks in RNA-seq data without the need for specific methodologies

Circulating ghrelin level is higher in HNF1A-MODY and GCK-MODY than in polygenic forms of diabetes mellitus

Ghrelin is a hormone that regulates appetite. It is likely to be involved in the pathophysiology of varying forms of diabetes. In animal studies, the ghrelin expression was regulated by the hepatocyte nuclear factor 1 alpha (HNF1A). Mutations of the HNF1A gene cause maturity onset diabetes of the young (MODY). We aimed to assess the circulating ghrelin levels in HNF1A–MODY and in other types of diabetes and to evaluate its association with HNF1A mutation status. Our cohort included 46 diabetic HNF1A gene mutation carriers, 55 type 2 diabetes (T2DM) subjects, 42 type 1 diabetes (T1DM) patients, and 31 glucokinase (GCK) gene mutation carriers with diabetes as well as 51 healthy controls. Plasma ghrelin concentration was measured using the immunoenzymatic assay with polyclonal antibody against the C-terminal fragment of its acylated and desacylated forms. Ghrelin concentrations were 0.75 ± 0.32, 0.70 ± 0.21, 0.50 ± 0.20, and 0.40 ± 0.16 ng/ml in patients with HNF1A–MODY, GCK–MODY, T1DM, and T2DM, respectively. The ghrelin levels were higher in HNF1A–MODY and GCK–MODY than in T1DM and T2DM (p < 0.001 for all comparisons) but lower than in non-diabetic controls (1.02 ± 0.29 ng/ml, p < 0.001 for both comparisons). In the multivariate linear model, the differences between both MODY groups and common diabetes types remained significant. Analysis by a HNF1A mutation type indicated that ghrelin concentration is similar in patients with different types of sequence differences. Plasma ghrelin level is higher in HNF1A–MODY and GCK–MODY than in the common polygenic forms of diabetes

The association of maternal gestational diabetes mellitus with autism spectrum disorders in the offspring

Wstęp. Istnieją dane wskazujące na związek między występowaniem cukrzycy przedciążowej u matki a ryzykiem zaburzeń ze spektrum autyzmu (ASD) u dziecka. Mniej jest dostępnych informacji dotyczących wpływu cukrzycy po raz pierwszy rozpoznanej w czasie ciąży (GDM) na ryzyko ASD. Badanie przeprowadzono w celu oceny częstości występowania ASD u dzieci matek, u których rozpoznano GDM. Materiał i metody. Autorzy przeanalizowali dokumentację medyczną pacjentek z GDM (947 kobiet, 1007 dzieci w wieku 4-6 lat) leczonych w Klinice Chorób Metabolicznych Szpitala Uniwersyteckiego w Krakowie w latach 1999-2011. Przeprowadzili również wywiad telefoniczny w celu zebrania danych klinicznych oraz informacji na temat parametrów biochemicznych. Wykonano test istotności, opierając się na rozkładzie dwumianowym prawdopodobieństwa w celu ustalenia, czy częstość występowania ASD u dzieci matek z GDM różniła się od dostępnych danych epidemiologicznych dotyczących ogólnej populacji polskich dzieci w wieku 0-18 lat. Sprawdzono również, czy istnieją inne istotne czynniki różnicujące matki z GDM w zależności od występowania ASD u ich potomstwa. Wyniki. Częstość występowania ASD u dzieci kobiet z GDM uczestniczących w badaniu autorów (8/1007) była wyższa niż w populacji polskich dzieci w wieku 0–18 (17,6/10 000; p = 0,0004). Stwierdzono, że w grupie matek dzieci z ASD przedciążowe mediany wartości wskaźnika BMI (20,862 vs. 23,529), SBP (110 mm Hg vs. 120 mm Hg) i DBP (70 mm Hg vs. 80 mm Hg) były niższe niż w grupie matek, u których dzieci nie rozpoznano ASD (odpowiednio p = 0,0349; p = 0,0149 i p = 0,0306). Urodzeniowa masa ciała dzieci z ASD była istotnie wyższa niż w przypadku dzieci bez ASD (3695 g vs. 3320 g; p = 0,0482). Wnioski. Częstość występowania ASD jest wyższa u dzieci matek z GDM niż w populacji ogólnej. Badanie potencjalnych czynników ryzyka ma podstawowe znaczenie dla lepszego zrozumienia tego zjawiska i ustalenia, jak mu zapobiec.Introduction. Some evidence exists for the association between exposure to pregestational maternal diabetes and risk of autism spectrum disorders (ASD) in offspring. Less information is available on the association of exposure to maternal gestational diabetes mellitus (GDM) with risk of ASD. We aimed to examine the prevalence of ASD disorders in offspring of mothers diagnosed with GDM. Material and methods. We analyzed data gathered from GDM patients (947 women; 1007 children aged 4-16 years) treated at the Department of Metabolic Diseases, University Hospital in Krakow from 1999 to 2011. We conducted a telephone survey to collect clinical information and biochemical parameters. We performed significance test based on the exact binomial probability to assess if the prevalence rate of ASD in offspring of mothers with GDM was different from available epidemiological data for children aged 0–18 years in Poland. We also checked whether there are any significant factors discriminating the mothers and offspring with and without ASD. Results. The prevalence of ASD in the offspring of mothers with GDM in our study (8/1007) was higher than in children aged 0-18 years in Poland (17.6/10000; p = 0.0004). The mothers of children with ASD had median pre-pregnancy BMI (20.862 vs. 23.529), SBP (110 mm Hg vs. 120 mm Hg) and DBP (70 mm Hg vs. 80 mm Hg) lower vs. group without ASD (p = 0.0349; p = 0.0149 and p = 0.0306 respectively). Birth weight of ASD children was significantly higher vs group without ASD (3695 g vs. 3320 g, p = 0.0482). Conclusions. The prevalence of ASD seems to be higher in offspring of mothers with GDM than in the general population. Studying the potential risk factors is crucial for better understanding of this phenomenon and it may be helpful to preventi it

DNA methylation analysis of negative pressure therapy effect in diabetic foot ulcers

Objective: Negative pressure wound therapy (NPWT) has been used to treat diabetic foot ulcerations (DFUs). Its action on the molecular level, however, is only partially understood. Some earlier data suggested NPWT may be mediated th rough modification of local gene expression. As methylation is a key epigenetic regulatory mechanism of gene expression, we assessed the effect of NPWT on its profile in patients with type 2 diabetes (T2DM) and neuropathic non-infected DFUs. Methods: Of 36 included patients, 23 were assigned to NPWT and 13 to standard therapy. Due to ethical concerns, the assignment was non-randomized and based on wound characteristics. Tissue samples were obtained before and 8 ± 1 days after therapy initiation. DNA methylation patterns were checked by Illumina Methylation EPIC kit. Results: In terms of clinical characteristics, the groups presented typical features of T2DM; however, the NPWT group had significantly greater wound ar ea: 16.8 cm2 vs 1.4 cm2 (P = 0.0003). Initially only one region at chromosome 5 was differentially methylated. After treatment, 57 differentially methylated genes were found, mainly located on chromosomes 6 (chr6p21) and 20 (chr20p13); they were associated with DNA repair and autocrine signaling via retinoic acid receptor. We performed differential analyses pre treatment and post treatment. The analysis reveale d 426 differentially methylated regions in the NPWT group, but none in the control group. The enrichment analysis showed 11 processes significantly associated with NPWT, of which 4 were linked with complement system activation. All but one were hypermethylated after NPWT. Conclusion: The NPWT effect on DFUs may be mediated through epigenetic chan ges resulting in the inhibition of complement system activation

Less but better : cardioprotective lipid profile of patients with GCK-MODY despite lower HDL cholesterol level

Patients with diabetes caused by single-gene mutations generally exhibit an altered course of diabetes. Those with mutations of the glucokinase gene (GCK-MODY) show good metabolic control and low risk of cardiovascular complications despite paradoxically lowered high-density lipoprotein (HDL) cholesterol levels. In order to investigate the matter, we analyzed the composition of low-density lipoprotein (LDL) and HDL subpopulations in such individuals. The LipoPrint(©) system (Quantimetrix, USA) based on non-denaturing, linear polyacrylamide gel electrophoresis was used to separate and measure LDL and HDL subclasses in fresh-frozen serum samples from patients with mutations of glucokinase or HNF1A, type 1 diabetes (T1DM) and healthy controls. Fresh serum samples from a total of 37 monogenic diabetes patients (21 from GCK-MODY and 16 from HNF1A-MODY), 22 T1DM patients and 15 healthy individuals were measured in this study. Concentrations of the small, highly atherogenic LDL subpopulation were similar among the compared groups. Large HDL percentage was significantly higher in GCK-MODY than in control (p = 0.0003), T1DM (p = 0.0006) and HNF1A-MODY groups (p = 0.0246). Patients with GCK-MODY were characterized by significantly lower intermediate HDL levels than controls (p = 0.0003) and T1DM (p = 0.0005). Small, potentially atherogenic HDL content differed significantly with the GCK-MODY group showing concentrations of that subfraction from control (p = 0.0096), T1DM (p = 0.0193) and HNF1A-MODY (p = 0.0057) groups. Within-group heterogeneity suggested the existence of potential gene–gene or gene–environment interactions. GCK-MODY is characterized by a strongly protective profile of HDL cholesterol subpopulations. A degree of heterogeneity within the groups suggests the existence of interactions with other genetic or clinical factors